The long-term objectives of this revised competing renewal are to provide data of therapeutic relevance to multiple sclerosis (MS) from neuroimmunologic studies on MS lesions and groups of interrelated experiments on the pathogenesis of immune-mediated demyelination. The proposed Program comprises three projects and a core facility and represents a request for funds to continue long-established, highly productive, collaborative research on MS. Dr. Raine will explore initiating events in CNS inflammation in MS and EAE from the standpoints of neuropathology, effector T cell characterization and adhesion molecule analysis (ICAM-1, LFA-1, VCAM-1, VLA-4). Novel aspects of this project will be a) the application of a congenic strain of mouse to adoptively-transferred EAE where donor T cells can be identified and separated from those of the recipient on the basis of a T cell marker (Thy-1.2) and b) a series of experiments to localize immune system molecules on surviving oligodendrocytes in acute MS lesions. Abrogation of EAE with reagents against adhesion molecules and cytokines, the quantitative expression of these molecules in treated animals, and a role for TCRgamma/delta T cells in autoimmune demyelination will also be investigated. Dr. Brosnan will examine interactions between proinflammatory cytokines (IL-1, TNFalpha) and members of the chemokine Beta family (e.g. MIP-1alpha and RANTES) on the development of the inflammatory response in the CNS. Different stages of disease activity will be analyzed for fluctuation in these factors and it will be determined whether they control specific subsets of T cells and macrophages and lead to macrophage activation in the lesions - information of possible therapeutic import for MS> Dr. Berman will explore two experimental systems of astrocyte/endothelial cell interactions with the goal of determining how the inflammatory response is controlled at the blood-brain barrier. The first is a co-culture system of human endothelial cells (EC) and syngeneic astrocytes. This model system will be used to examine how coculture of EC and astrocytes affects their response to inflammatory mediators. Functional changes will be determined by leukocyte adhesion and migration assays. The second system will study cytokine-induced inflammation at different sites in the rabbit CNS using a combination of pathologic and electrophysiologic approaches. In this system, the question of regional specialization in the response of the CNS vasculature to these inflammatory agents will be addressed. Since MS might represent a collection of CNS disorders, the aims of this program might provide new criteria for the dissection of its various forms and disease stages.